Medical Research Council National Institute for Medical Research, Division of Molecular Neurobiology, London, UK.
Nat Methods. 2011 Mar;8(3):260-6. doi: 10.1038/nmeth.1567. Epub 2011 Feb 6.
To facilitate studies of neural network architecture and formation, we generated three Drosophila melanogaster variants of the mouse Brainbow-2 system, called Flybow. Sequences encoding different membrane-tethered fluorescent proteins were arranged in pairs within cassettes flanked by recombination sites. Flybow combines the Gal4-upstream activating sequence binary system to regulate transgene expression and an inducible modified Flp-FRT system to drive inversions and excisions of cassettes. This provides spatial and temporal control over the stochastic expression of one of two or four reporters within one sample. Using the visual system, the embryonic nervous system and the wing imaginal disc, we show that Flybow in conjunction with specific Gal4 drivers can be used to visualize cell morphology with high resolution. Finally, we demonstrate that this labeling approach is compatible with available Flp-FRT-based techniques, such as mosaic analysis with a repressible cell marker; this could further support the genetic analysis of neural circuit assembly and function.
为了促进神经网络结构和形成的研究，我们生成了三种果蝇（Drosophila melanogaster）变体的小鼠 Brainbow-2 系统，称为 Flybow。编码不同膜固定荧光蛋白的序列被排列在侧翼带有重组位点的盒内对中。Flybow 结合 Gal4-上游激活序列二元系统来调节转基因的表达，并结合诱导型修饰的 Flp-FRT 系统来驱动盒的倒位和切除。这为一个样本中两个或四个报告基因中的一个的随机表达提供了空间和时间上的控制。通过使用视觉系统、胚胎神经系统和翅 imaginal 盘，我们表明，Flybow 与特定的 Gal4 驱动子结合使用可以以高分辨率可视化细胞形态。最后，我们证明这种标记方法与现有的基于 Flp-FRT 的技术兼容，例如带有可抑制细胞标记物的马赛克分析；这可以进一步支持神经回路组装和功能的遗传分析。
